Mineralocorticoid receptor mediates vascular stiffness via dysregulated immunity in perivascular adipose tissue

项目总结 我的研究集中在血管周围的盐皮质激素受体(Mr)信号之间的相互作用。 脂肪组织(PVAT)、免疫失调和血管功能障碍在高脂高脂饮食中的作用 果糖饮食(西方饮食[WD])WD会导致胰岛素抵抗。重要的是，胰岛素抵抗会导致 女性患有侵袭性心血管疾病。我假设在女性中，WD诱导的胰岛素抵抗 导致T调节(Treg)功能丧失，导致巨噬细胞MR激活和随后的M1 PVAT中的极化。此外，我假设这些事件通过激活 组织转谷氨酰胺酶-2(TG2)部分通过氧化应激和生物可利用的一氧化氮减少 (否)。为了验证我的假设，具体目标1是确定磁共振激活PVAT巨噬细胞的效果 血管僵硬的发病机制及其与Treg功能受损和M1巨噬细胞的关系 极化。髓系特异性MR基因敲除(MyMRKO)雄性和雌性小鼠将接受WD或 醛固酮和我们将测量主动脉硬度。在PVAT中，我们将测量巨噬细胞极化和 发炎。我们将测量原代内皮细胞(EC)和血管平滑肌的细胞硬度 在PVAT条件培养液中培养的细胞(VSMC)来自每个队列。在具体目标2中，我们将确定 Tregs对PVAT巨噬细胞极化和主动脉僵硬的作用。在这组实验中，我们将使用 雄性和雌性C57BL/6J小鼠过继Treg转移给WD或WD或WD组 缓慢加压剂量的醛固酮。我们将测量巨噬细胞极化、炎症和主动脉 硬度，以及在PVAT条件下的介质中的EC/VSMC硬度测量。我已经组建了一支队伍 导师和合作者将在整个项目中指导我，并促进我作为 独立研究人员。我的职业发展计划有一个初始训练期(1-3年)，在这段时间里我将 完成拟议的工作，并有一个过渡期(4-5年)，在此期间我将专注于R01提案。 我的指导团队拥有必要的基础设施、专业知识和资源，我的机构将提供 必要的支持使我能够成功地实现我的目标。我的中间目标是成功地 完成我提议的工作。我的最终目标是建立一条明确的工作路线，解决这些机制 MR激活和免疫失调可导致血管疾病。

Project summary My research is focused on the interactions between mineralocorticoid receptor (MR) signaling in perivascular adipose tissue (PVAT), dysregulation of immunity and vascular dysfunction in the setting of high-fat high- fructose diet (Western Diet [WD]). WD leads to insulin resistance. Importantly, insulin resistance leads to aggressive cardiovascular disease in females. I hypothesize that in females, WD-induced insulin resistance results in loss of T regulatory (Treg) function leading to macrophage MR activation and subsequent M1 polarization in PVAT. Further, I hypothesize that these events increase vascular stiffness via activation of Tissue transglutaminase-2 (TG2), in part, through oxidative stress and decreased bioavailable nitric oxide (NO). To test my hypothesis, specific Aim 1 is to determine the effect of MR activation in PVAT macrophages on the pathogenesis of vascular stiffness as it relates to impaired Treg function and M1 macrophage polarization. Myeloid-specific MR knockout (MyMRKO) male and female mice will be treated with a WD or aldosterone and we will measure aortic stiffness. In PVAT, we will measure macrophage polarization and inflammation. We will measure cellular stiffness in primary endothelial cells (EC) and vascular smooth muscle cells (VSMC) cultured in PVAT-conditioned media from each cohort. In Specific Aim 2, we will determine the role of Tregs on PVAT macrophage polarization and aortic stiffness. In this set of experiments, we will use adoptive Treg transfer from male and female C57Bl/6J mice fed a normal diet to the mice treated with WD or slow pressor doses of aldosterone. We will measure macrophage polarization, inflammation and aortic stiffness, along with EC/VSMC stiffness measurement in PVAT-conditioned media. I have assembled a team of mentors and collaborators that will guide me throughout this project and promote my transition as an independent researcher. My career development plan has an initial training period (years 1-3) in which I will complete the proposed work, and a transitional period (years 4-5) during which I will focus on an R01 proposal. My mentoring team has the necessary infrastructure, expertise and resources and my institution will provide the necessary support to enable me successfully achieve my goals. My intermediate goal is to successfully complete the work I propose. My ultimate goal is to establish a distinct line of work addressing the mechanisms by which MR activation and dysregulated immunity lead to vascular disease.